Silver Nanoparticles as an Electrochemical Bio-Label

Paul K. Wilson1, Mateusz Szymanski2, Robert A. Porter1* 1. Argento Diagnostics Ltd., Knowledge & Innovation Centre, National Physical Laboratory, Teddington, Middlesex, TW11 0LW 2. Analytical Science Team, National Physical Laboratory, Teddington, Middlesex, TW11 0LW

Introduction
Immunoassays with electrochemical detection of metal marker generally follows the same procedure. That is the dissolution of metal nanoparticle (NP) and the consecutive quantification of solubilized metal ions by anodic striping voltammetry (ASV) at carbon electrode. The dissolution step is very important as it ensures the high sensitivity due to release of huge number of metal ions from one single NP. However, it requires the use of harmful reagents (such as HBr/Br2 mixture for gold or HNO3 for silver). This work presents a new strategy for using silver nanoparticles (AgNPs) as a metal marker with the dissolution step that does not require a chemical oxidant and thus offering advantages for application in terms of fast response, reduced cost of analysis and procedural simplicity.

Electrochemical behaviour of AgNPs

Electrochemical reactions at the working electrode. 1. Silver sol dissolution and formation of electroactive chelate: amonium thiocyanatosilver(I) at +0.6 V for 15 s.

Ag
Ag

Ag
4

e
n ( SCN
n 1

m ( NH

[ Ag ( NH

) m ( SCN

) n ]m

n 1

2. Accumulation step of ASV (reduction) at 1.2V

[ Ag ( NH 4 ) m ( SCN ) n ] m

[ Ag ( NH 4 ) m ( SCN ) n ] m
+0.1V
n 1

3. Stripping step of ASV (oxidation) -1.2V

[ Ag ( NH 4 ) m ( SCN ) n ]

m n

[ Ag ( NH 4 ) m ( SCN ) n ] m

Silver Nanoparticles Aggregation

AgNPs in the presence of thiocyanate forms aggregates surrounded with the SCN- ion layer.

The analytical signal of different concentration of Silver colloid.

ASV scans of silver colloid (4-80%) in 1M NH4SCN

0.201x10-4 0.176x10-4 0.151x10-4 0.126x10-4 i/A
Microcoulombs [C] 3 2.5 2 1.5 1 0.5 0 0 20 40 Silver Sol [% ] 60 80 y = 0.0305x - 0.028 2 R = 0.9993

2.500x10 0

-1

Silver Sol in 1M NH4SCN

-2.500x10 -5.000x10 -7.500x10 -10.000x10 -12.500x10 -15.000x10 61.783 62.033 62.283 t/s 62.533 62.783 -17.500x10

-1

SCN-

SCNSCN-

-1

0.101x10-4 0.076x10-4 0.051x10-4

E/V

-1

NH4SCN

SCNSCNSCNSCN-

-1

SCNSCNSCN-

0.026x10

-4

-1

0.001x10-4 -0.024x10-4 -0.049x10-4 61.533

-1

-1

= 40 nm Zeta Potential = -50 mV

= 400 nm Zeta Potential = -10 mV Mechanism of AgNPs dissolution.

Both UV-Vis and Dynamic Light Scattering confirm the aggregation process.

Carbon Screen Printed Electrodes

The measurement takes place in 50-L droplet on the surface of a carbon screen printed electrode.
Counter Electrode Working Electrode

The influence of dissolution step parameters on ASV signal.

NH4SCN molarity
Reference Electrode
10

Potential value
9

Duration
10

10

2.5 ppm Ag+ 80% Silver Sol

M ic ro c o u lo m b s [C ]

8 6 4

6
6 1 5

2 0 0 0.5

Steps of Anodic Stripping Voltammetry:

1 [NH4SCN] [mol/L]

1.5

3 -1.6

-1.1

-0.6

-0.1

0.4

0.9

1.4

0 0 10 time [s] 20 30 40

potential [V]

AgNPs dissolution +0.6 V Stripping (oxidation)

Conclusion
We have demonstrated and explained the mechanism of AgNPs dissolution on the surface of carbon electrode without the need of any oxidant. This process can be applied for development of electrochemical metalloimmunoassays. Removing the need for using harsh oxidant is very advantageous especially in terms of building new point of care tests.
This project was funding by the UK Governments Chemistry and Biology National Measurement System Programme under the Bioanalysis theme. * Contact: robert.porter@argentodiagnostics.com

plating (reduction) -1.2 V -1.6 V nucleation

Time

[s]